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Results: 7

1.
Figure 6.

Figure 6. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

In vivo association of CD30v protein with TRAF2 and TRAF5. Cell lysates of 293T cells transfected with expression vectors indicated above the figure were immunoprecipitated with HCD30C2 or anti-CD30 mAb Ber-H2, and immune complexes were analyzed by immunoblotting with anti-FLAG M2 antibody (top). Positions of the FLAG-tagged TRAF2 and TRAF5 are indicated on the right, and those of molecular weight markers on the left. Expression of FLAG-tagged TRAF2 and TRAF5 was detected by M2 antibody, using whole cell lysates (*, bottom).

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
2.
Figure 5.

Figure 5. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

Correlation between expressions of CD30v and CD30L. Expression of transcripts for CD30v and CD30L, as detected by RT-PCR in myeloid (square) and lymphoid (circle) hematopoietic malignancies was compared. Figures in squares or circles represent numbers of cases, within the whole group of myeloid (squares) or lymphoid (circles) hematopoietic malignancies, expressing a given combination of CD30v and CD30L mRNAs. Sums are reported as unframed figures for each combination of CD30v and CD30L mRNA expression. The χ 2 test used for these latter figures revealed a P value of 0.02 and an odds ratio of 3.22.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
3.
Figure 2.

Figure 2. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

Specificity of the peptide antibody. A: Immunoblot analysis of CD30v. a: Immunoblotting with Ber-H2 detected CD30 proteins (arrows) but not CD30v (arrowhead). An asterisk indicates the position of the immunoglobulin heavy chain. The lane labeled “vector” indicates a sample prepared from 293T cells transfected with an empty vector (pME18S). b: HCD30C2 reacted with CD30v proteins (arrow) but not with CD30 proteins. An asterisk to the right of the figure indicates the position of a nonspecific band detected in all samples. Arrowheads indicate the positions of immunoglobulins. B: Competition of CD30v immunoprecipitation by the immunogen peptide. Immunoprecipitation of 32P-labeled 293T cells transfected with CD30v expression vector by HCD30C2, with or without pretreatment with the immunogen (10 μg), as indicated above the figure. Competitor + indicates pretreatment of HCD30C2 with the immunogen.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
4.
Figure 3.

Figure 3. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

CD30v protein in cells detected with the use of a specific antibody. A: Indirect immunofluorescence cytochemistry of transfected cell lines. COS-7 cells transfected with pME-CD30v (a and b) or pME-CD30 (c and d) were examined using HCD30C2 (a and c) or Ber-H2 (b and d). B: Indirect immunofluorescence cytochemistry of HL-60 and AML samples. Cells were examined using HCD30C2 antibody. a, untreated HL-60; b, TPA-treated HL-60; c and d, AML samples. C: Immunohistochemistry of formalin-fixed, paraffin-embedded bone marrow samples. CD30v protein was evident in leukemia cells but not in normal bone marrow samples. a–c, bone marrow samples of leukemia patients (AML M3, AML M4, and ALL L2, respectively); d, a normal bone marrow sample.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
5.
Figure 7.

Figure 7. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

RT-PCR analysis of transcripts for TRAF1, 2, 3, and 5 in leukemic blasts. CD30v-expressing samples were used for the study. After 40 cycles of PCR in a 50-μl reaction volume, a 10-μl aliquot was analyzed by agarose gel electrophoresis and ethidium bromide staining. Among the TRAF proteins, those reported to interact with CD30 were examined. The expected size for each PCR product is indicated on the right. a: CD30v; b: TRAF1; c: TRAF2; d: TRAF3; e: TRAF5; M2, M4, M5, FAB classification. ATL, adult T-cell leukemia; HL-60/TPA, TPA-treated HL-60; H2O, PCR without template cDNA; m, molecular weight marker.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
6.
Figure 4.

Figure 4. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

Combined analysis of CD30v expression with Northern blot and immunoblot. A: Expression of CD30v gene in hematopoietic neoplasms. Total RNA (10 μg/lane) from AML (M1, M2, M4, and M5 phenotype) and HCL (two cases) was analyzed by Northern blotting with a CD30 probe. Untreated or TPA-treated HL-60 cells served as negative and positive controls for CD30v expression, respectively. The Karpas 299 cell line, derived from a CD30+ large anaplastic lymphoma, was used as a positive control for CD30 expression. Positions for CD30 and CD30v mRNAs are indicated on the right. The same blot was also probed using a specific β-actin cDNA fragment (bottom). B: Detection of CD30v protein in cell lysates from three AML (M1, M4, and M5) and two HCL cases by immunoblot with the HCD30C2 antiserum. Positions for CD30v protein and molecular weight markers are indicated on the far right and left of the figure, respectively.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.
7.
Figure 1.

Figure 1. From: Frequent Expression of the Variant CD30 in Human Malignant Myeloid and Lymphoid Neoplasms.

RT-PCR analysis of CD30v transcripts. A: Position of the primer pairs for CD30v and CD30. Common sequences between CD30 and CD30v cDNA are indicated by the bold line. Unique sequences in the CD30v cDNA are indicated by the dotted line. Paired arrows A, B, and C indicate positions of each primer in the sequences. B: Specific amplification of the cDNA template by CD30v primers. PCR was run for 40 cycles in a 50-μl reaction volume, and a 10-μl aliquot was analyzed by agarose gel electrophoresis and ethidium bromide staining. Lanes 1 and 2, cDNA samples prepared from RNA, without or with DNase treatment, respectively; lane 3, an RNA sample without reverse transcription; lane 4, a control genomic DNA sample; lane 5, without template. m, molecular weight marker. C: The primer pair for CD30v does not cross-amplify CD30 transcripts. RT-PCR analysis of CD30v (top) and CD30 (bottom) in the CD30-positive cell lines: L428, KMH-2, HUT-102, and K562. m, molecular weight marker. D and E: RT-PCR analysis of CD30v transcripts in myeloid leukemia samples (D) and samples of LPD (E). Representative results of Southern blot analysis of RT-PCR products are shown in the top panels. Probes used for hybridization are indicated on the left. The bottom panel shows the ethidium bromide staining of the agarose gel. Positions for PCR products corresponding to the longer and shorter CD30v transcripts (CD30vs and CD30vl, respectively) are indicated on the right. M0, M1, M2, M3, M4, and M5 indicate FAB classification. ALL, acute lymphoblastic leukemia; CLL, chronic lymphocytic leukemia; PLL, prolymphocytic leukemia; HCL, hairy cell leukemia; NHL, non-Hodgkin’s lymphoma; HG, high grade; LG, low grade; MM, multiple myeloma; ATL, adult T-cell leukemia; (+), positive control; (−), negative control.

Ryouichi Horie, et al. Am J Pathol. 1999 December;155(6):2029-2041.

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